Human Alveolar Epithelial Cells Expressing Tight Junctions to Model the Air-Blood Barrier
Human Alveolar Epithelial Cells Expressing Tight Junctions to Model the Air-Blood Barrier
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DOI：
10.14573/altex.1511131
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发表时间：
2016-01-01
影响因子：
5.6
通讯作者：
Lehr, Claus-Michael
中科院分区：
文献类型：
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作者：
Kuehn, Anna;Kletting, Stephanie;Lehr, Claus-Michael
本文描述了一种新的人肺泡上皮细胞系（hAELVi -人肺泡上皮慢病毒永生化），具有I型样特征和功能性紧密连接，适用于模拟外周肺的空气-血液屏障。原代人肺泡上皮细胞永生化的一种新的方案，生长为单层的可渗透的过滤器支持，其特征在于形态学，生化和生物病理学。hAELVi细胞保持形成紧密细胞间连接的能力，具有高跨上皮电阻（> 1000 Ω（星星）cm（2））。在液-液以及气-液条件下，细胞可保持培养数天，直至第75代。超微结构分析和实时PCR显示I型样细胞的特性，如小窝的存在，小窝蛋白-1的表达，和缺乏表面活性蛋白C。考虑到屏障性质，还观察到用紧密连接和桥粒密封的相互指状结构。亲水性标记物荧光素钠的低渗透性证实了hAELVi细胞适用于跨肺泡上皮的体外转运研究。这些结果表明，hAELVi细胞反映了空气-血液屏障的基本特征，这是研究吸入药物、化学品和纳米材料的吸收和毒性的动物试验的替代方法所需要的。
This paper describes a new human alveolar epithelial cell line (hAELVi - human Alveolar Epithelial Lentivirus immortalized) with type I-like characteristics and functional tight junctions, suitable to model the air-blood barrier of the peripheral lung. Primary human alveolar epithelial cells were immortalized by a novel regimen, grown as monolayers on permeable filter supports and characterized morphologically, biochemically and biophysically. hAELVi cells maintain the capacity to form tight intercellular junctions, with high trans-epithelial electrical resistance (> 1000 Omega(star)cm(2)). The cells could be kept in culture over several days, up to passage 75, under liquid-liquid as well as air-liquid conditions. Ultrastructural analysis and real-time PCR revealed type I-like cell properties, such as the presence of caveolae, expression of caveolin-1, and absence of surfactant protein C. Accounting for the barrier properties, inter-digitations sealed with tight junctions and desmosomes were also observed. Low permeability of the hydrophilic marker sodium fluorescein confirmed the suitability of hAELVi cells for in vitro transport studies across the alveolar epithelium. These results suggest that hAELVi cells reflect the essential features of the air-blood barrier, as needed for an alternative to animal testing to study absorption and toxicity of inhaled drugs, chemicals and nanomaterials.